Figure 4

Figure 4

Molecular evidence for the hyperactivation of mTOR signaling in PIKfyve mutant zebrafish. (A) Western blot analysis showing the sustained mTOR activity during early development in PIKfyve mutant zebrafish embryos. (B) Densitometric analysis of phosphorylation status of 4E-BP1 and S6K in (A) (ns, non-significant; * p < 0.05, ** p < 0.01; two-way ANOVA). (C) Western blot analysis of mTOR activity in WT and MU zebrafish embryos at 5 dpf with or without rapamycin treatment. (D) Densitometric analysis of phosphorylation status of 4E-BP1 and S6K in (C) (ns, non-significant; ** p < 0.01, *** p < 0.001, **** p < 0.0001; two-way ANOVA). (E) WT and MU zebrafish embryos with or without rapamycin treatment were fixed at 5 dpf, permeabilized, and immuno-stained with phospho-S6K antibody. Arrows denote the macrophages with intense phospho-S6K staining. Scale bar, 20 µm.